Anti neun primary antibody

Primary cortical cultures prepared from El8 fetal Sprague-Dawley rat brains were cultured on coverslips in 24-well plates. After 2 days of Aβ_25-35 incubation, the cell culture medium was removed and replaced with BrdU labeling solution and incubated at 37°C for 4 days along with LiBen (0.5 mM), LiCl (0.5 mM), or SB (0.5 mM) treatments. Subsequently, cells were fixed in 4% paraformaldehyde solution and washed with PBS, blocked with 2% bovine serum albumin, permeabilized with 0.03% Triton X-100 in PBS, and acid-washed with 2 N HCl and phosphate/citric acid buffer (pH 7.4). Next, anti-BrdU primary antibody (1 : 250, Thermo Fisher Scientific) or anti-NeuN primary antibody (1 : 300, Abcam) were added. The goat anti-rat IgG Alexa Fluor^® 488-conjugated secondary antibodies (1 : 200; Cat. No. A11006; Thermo Fisher Scientific); anti-mouse IgG Alexa Fluor^® 546-conjugated secondary antibodies (1 : 500; Cat. No. A11003; Thermo Fisher Scientific) were applied to bind the primary antibodies of the BrdU and NeuN respectively. The coverslips were observed under a digital imaging fluorescence microscope (Olympus BX61, Japan) equipped with filter sets to detect the corresponding fluorescence signals.